Comparative Progression of Comparative Progression of Atherosclerosis in ApoE-/- Mice Atherosclerosis in ApoE-/- Mice

Exposed to ETS and CAPsExposed to ETS and CAPs

Lung Chi Chen, Ph. D.Lung Chi Chen, Ph. D.Department of Environmental MedicineDepartment of Environmental Medicine

NYU School of MedicineNYU School of Medicine

7th ICCAD, Venice Italy7th ICCAD, Venice ItalyOct. 10, 2007Oct. 10, 2007

SCHOOL OFMEDICINE

NEW YORK UNIVERSITY

Health Risks of Environmental Health Risks of Environmental Tobacco Smoke (ETS) ExposureTobacco Smoke (ETS) Exposure

ETS increases coronary heart disease (CHD) ETS increases coronary heart disease (CHD) in non-smokers by a factor of 1.3.in non-smokers by a factor of 1.3.

Long term exposure to ETS caused 40,000 - Long term exposure to ETS caused 40,000 - 60,000 excess heart disease deaths in the 60,000 excess heart disease deaths in the US.US.

Smoking produces atherogenic serum lipid Smoking produces atherogenic serum lipid profileprofile

Smoking increases intima-media thickness Smoking increases intima-media thickness coronary progressioncoronary progression

When Pollution Gets High, Deaths Increase:When Pollution Gets High, Deaths Increase:1952 London Fog Episode Deaths vs. PM1952 London Fog Episode Deaths vs. PM

(Pope, Burnett, Thun, Calle, Krewski, Ito , and Thurston) (Pope, Burnett, Thun, Calle, Krewski, Ito , and Thurston) ((JAMAJAMA, 2002), 2002)

Association between Ambient Particle Pollution and Atherosclerosis

Künzli et al, EHP 113, 201-206, 2005In Los AngelesCIMT: carotid intima-media thickness

Schematic of the evolution of the Schematic of the evolution of the atherosclerotic plaqueatherosclerotic plaque

Braunwald's Heart Disease: A Textbook of Cardiovascular Medicine, 7th ed

Common Mechanisms of Ambient Particles and Tobacco Smoke Induced Atherosclerosis

Ambient ParticlesCigarette Smoke

Oxidative Stress(O2

-, H2O2, ONOO-)

Vascular Dysfunction, Prothrombotic Factors,Leukocyte, Platelet Activation, Lipid PeroxidationAdhesion Molecules, Smooth Muscle Proliferation

Initiation and Progression ofAtherothrombotic Diseases

InflammatoryGene Activation

NO Regulation

EXPERIMENTAL DESIGNEXPERIMENTAL DESIGNEXPERIMENTAL DESIGNEXPERIMENTAL DESIGN ApoEApoE-/--/- mice mice

Normal Chow (Normal Chow (4% fat, w/w, 8% calories from fat)4% fat, w/w, 8% calories from fat) High Fat Chow (High Fat Chow (21% fat, w/w, 42% calories from fat)21% fat, w/w, 42% calories from fat)

Exposed, whole body, to fine CAPs or filtered air for 6 hours/day, 5 days per Exposed, whole body, to fine CAPs or filtered air for 6 hours/day, 5 days per weekweek May 29, - Dec. 7, 2007.May 29, - Dec. 7, 2007. Average 6 hr Concentration: 165 µg/mAverage 6 hr Concentration: 165 µg/m33

equivalent to 30 ug/mequivalent to 30 ug/m33 annual average. annual average. Exposed, whole body, to side stream (SS) smoke or filtered air for 6 hours/day, Exposed, whole body, to side stream (SS) smoke or filtered air for 6 hours/day,

5 days per week, for 6 months5 days per week, for 6 months Average 6 hr Concentration: 450 µg/mAverage 6 hr Concentration: 450 µg/m33

Approximately equivalent to living with a smoker who smoke 3 cigarettes/day.Approximately equivalent to living with a smoker who smoke 3 cigarettes/day.

Ambient Particle Concentrating and Exposure SystemsAmbient Particle Concentrating and Exposure SystemsAmbient Particle Concentrating and Exposure SystemsAmbient Particle Concentrating and Exposure Systems

Animal Exposure Chambers

VACES

Modified DataScience ECG Receivers

Inside of the exposure chambers

ETS ExposureETS ExposureSide-Stream Smoking MachineSide-Stream Smoking Machine

(CSM 2070, CH Tech)(CSM 2070, CH Tech)

2R4F, 2 sec 35 ml puff/min, 12 puffs/cigarette2R4F, 2 sec 35 ml puff/min, 12 puffs/cigarette Whole-body exposure chamberWhole-body exposure chamber

EXPERIMENTAL DESIGNEXPERIMENTAL DESIGNEXPERIMENTAL DESIGNEXPERIMENTAL DESIGN

ECG tracing monitored continuously for daily ECG tracing monitored continuously for daily changes in heart rate (HR) and heart rate changes in heart rate (HR) and heart rate variability (HRV)variability (HRV)

At 2, 4, 6 months of ETS exposure or at 3 and 6 At 2, 4, 6 months of ETS exposure or at 3 and 6 months of CAPs exposuremonths of CAPs exposure

Ultrasound Biomicroscopy (UBM)Ultrasound Biomicroscopy (UBM)

Vascular ReactivityVascular Reactivity

HistopathologyHistopathology

H&EH&E

ImmunohistostainingImmunohistostaining

Ultrasound Biomicroscopy (UBM)Ultrasound Biomicroscopy (UBM)Ultrasound Biomicroscopy (UBM)Ultrasound Biomicroscopy (UBM)

Vevo 660, VisualSonics, Toronto, OntarioVevo 660, VisualSonics, Toronto, Ontario single-crystal mechanical transducersingle-crystal mechanical transducer central frequency: 40 MHz central frequency: 40 MHz focal length: 6 mmfocal length: 6 mm frame rate: 30 Hzframe rate: 30 Hz maximum field of view of 2D imaging: 10x10 mmmaximum field of view of 2D imaging: 10x10 mm spatial resolution: spatial resolution: ~60 µm (lateral) by ~30 µm (axial) ~60 µm (lateral) by ~30 µm (axial) Doppler pulse repetition frequency: up to 96 kHz, maximum Doppler pulse repetition frequency: up to 96 kHz, maximum

unaliased velocity of 120 cm/sunaliased velocity of 120 cm/s

Mice were anesthetized using 1.5% isofluraneMice were anesthetized using 1.5% isoflurane Total scan time: 30 min/mouseTotal scan time: 30 min/mouse Advantage:Advantage: Non-invasive, Progression Non-invasive, Progression

Ultrasound Scan ProcedureUltrasound Scan Procedure

Anesthetized: continuous 1.5% isoflurane

Total scan time: 30min/mice

Ultrasound Images of Aorta Plaques

Ascending Aorta Aorta Arch

LeftCommonCorotidartery

Brachio-Cephalicartery

RightCommonCorotidartery

UBM Quantification ProcedureUBM Quantification Procedure

330um

Atherosclerosis Progression inNaïve ApoE-/-

At baseline, Young: 1.5 month old; Old: 10.5 month old

0

10

20

30

40

50

Percent Plaque Volume

Baseline 2-month4-month6-month9-month

Young NCYoung HFC

Old NCOld HFC

Left Common Carotid Artery

Young NCYoung HFCOld NCOld HFC

Longevity of the ApoELongevity of the ApoE-/--/- mice mice reduced by SS but not by CAPsreduced by SS but not by CAPs

No death occurred in ApoE-/- mice in CAPs exposed group.

CAPs Study in 2003

SS Enhanced Atherogenesis (UBM)

4 Month--Brachiocephalic Artery

Air ETS0

5

10

15

20

25

30

35

Exposure

4 Month--Carotid Artery

Air ETS0

5

10

15

20

25

30

35

Exposure

6 Month -- Brachiocephalic Artery

Air ETS0

5

10

15

20

25

30

35

Exposure

6 Month--Carotid Artery

Air ETS0

5

10

15

20

25

30

35

Exposure

Normal Chow

High Fat Chow

*

SS Enhanced Atherogenesis(within the same animals)

CAPs exposure enhanced atherogenesis in ApoE-/-

mice, with accompanied increases in lipid content

H&EH&E

OROORO

AirAir CAPsCAPs AirAir CAPsCAPs

Normal ChowNormal Chow High Fat ChowHigh Fat ChowSun et al, JAMA, 2005

Comparative Atherogenic Effects of Comparative Atherogenic Effects of CAPs vs SS (UBM)CAPs vs SS (UBM)

0

20

40

60

80

100

Percentage Increase

Over Control

Normal Chow High Fat Chow

SSCAPs (85 µg/m3)CAPs (165µg/m3)

CAPs: Aortic Arch and Bachiocephalic Artery; SS: Carotid Artery

SummarySummary UBM is a useful imaging technique to follow lesion UBM is a useful imaging technique to follow lesion

progression in atherosclerotic mice.progression in atherosclerotic mice.

Carotid artery appears to be the optimal location to Carotid artery appears to be the optimal location to evaluate atherosclerosis progression.evaluate atherosclerosis progression.

SS, but not CAPs, reduced longevity in ApoESS, but not CAPs, reduced longevity in ApoE-/--/-..

CAPs and SS exposures promote similar magnitude of CAPs and SS exposures promote similar magnitude of change in animals susceptible to atherosclerosis, change in animals susceptible to atherosclerosis, regardless of the diet. regardless of the diet.

Exacerbation of atherosclerosis is evident after 4 months Exacerbation of atherosclerosis is evident after 4 months of low level SS exposure or after 3 months of CAPs of low level SS exposure or after 3 months of CAPs exposure.exposure.

Acknowledgements:

Qinghua Sun (Ohio State University) Sanjay Rajagopalan (Ohio State University) Chunli Quan Mianhua Zhong Ximei Jin Qiang Li

Thank You !!!Thank You !!!

Contact Information:Tel: (845) 731-3560

E-mail: LCC4@nyu.edu